Articles by Gary Zenitsky in JoVE
Rapid and Refined CD11b Magnetic Isolation of Primary Microglia with Enhanced Purity and Versatility Souvarish Sarkar*1, Emir Malovic*1, Brandon Plante1, Gary Zenitsky1, Huajun Jin1, Vellareddy Anantharam1, Arthi Kanthasamy1, Anumantha G. Kanthasamy1 1Biomedical Sciences & Iowa Center for Advanced Neurotoxicology, Iowa State University Here, we present a protocol to isolate microglia from postnatal mouse pups (day 1) for in vitro experimentation. This improvised method of isolation generates both high yield and purity, a significant advantage over alternate methods that allows broad range experimentation for the purposes of elucidating microglial biology.
Other articles by Gary Zenitsky on PubMed
Inactivation of the Brachium Conjunctivum Prevents Extinction of Classically Conditioned Eyeblinks Brain Research. May, 2005 | Pubmed ID: 15910776 It is well established that the intermediate cerebellum is involved in the acquisition of classically conditioned eyeblink responses (CRs). Recent studies that inactivated the interposed nuclei (IN) demonstrated that blocking the intermediate cerebellum also interrupts CR extinction. Is this extinction deficit related to interrupting the information flow to efferent targets of the IN? To address this question, we inactivated axons of IN neurons in the brachium conjunctivum (BC). This treatment blocked the output of the intermediate cerebellum without directly affecting neurons in the deep cerebellar nuclei. Rabbits were trained in a delay classical conditioning paradigm, using a tone as the conditioned stimulus (CS) and a corneal air puff as the unconditioned stimulus (US). Then, the BC was microinjected with a sodium channel blocker, tetrodotoxin, during 4 extinction sessions in which rabbits were presented only with the CS. Tests performed after the 4-day injection period revealed that CRs did not extinguish in BC inactivation sessions but extinguished at a normal rate in the absence of the drug. CRs were then re-acquired. These data show that the normal flow of information along axons of cerebellar nuclear cells is required for CR extinction.
A Trigeminal Conditioned Stimulus Yields Fast Acquisition of Cerebellum-dependent Conditioned Eyeblinks Behavioural Brain Research. Jan, 2012 | Pubmed ID: 21933685 Classical conditioning of the eyeblink response in the rabbit is a form of motor learning whereby the animal learns to respond to an initially irrelevant conditioned stimulus (CS). It is thought that acquired conditioned responses (CRs) are adaptive because they protect the eye in anticipation of potentially harmful events. This protective mechanism is surprisingly inefficient because the acquisition of CRs requires extensive training - a condition that is unlikely to occur in nature. We hypothesized that the rate of conditioning in rabbits could depend on CS modality and that stimulating mystacial vibrissae as the CS could produce CR acquisition faster than the traditional auditory or visual stimulation. We tested this hypothesis by conditioning naïve rabbits in the delay paradigm using a weak airpuff CS (vCS) directed to the ipsilateral mystacial vibrissae. We found that the trigeminal vCS yields significantly faster CR acquisition. We next examined if vCS-evoked CRs are dependent on the intermediate cerebellum in the same fashion as CRs evoked by the traditional auditory CS. We found that vibrissal CRs could be abolished by inactivating the cerebellar interposed nuclei (IN) with muscimol. In addition, injections of picrotoxin in the IN shortened the onset latency of vibrissal CRs. These findings suggest that the tone and vCS-evoked CRs share similar cerebellar dependency.
Blocking Glutamate-mediated Inferior Olivary Signals Abolishes Expression of Conditioned Eyeblinks but Does Not Prevent Their Acquisition The Journal of Neuroscience : the Official Journal of the Society for Neuroscience. May, 2013 | Pubmed ID: 23699520 The inferior olive (IO) is considered a crucial component of the eyeblink conditioning network. The cerebellar learning hypothesis proposes that the IO provides the cerebellum with a teaching signal that is required for the acquisition and maintenance of conditioned eyeblinks. Supporting this concept, previous experiments showed that lesions or inactivation of the IO blocked CR acquisition. However, these studies were not conclusive. The drawback of the methods used by those studies is that they not only blocked task-related signals, but also completely shut down the spontaneous activity within the IO, which affects the rest of the eyeblink circuits in a nonspecific manner. We hypothesized that more selective blocking of task-related IO signals could be achieved by using injections of glutamate antagonists, which reduce, but do not eliminate, the spontaneous activity in the IO. We expected that if glutamate-mediated IO signals are required for learning, then blocking these signals during training sessions should prevent conditioned response (CR) acquisition. To test this prediction, rabbits were trained to acquire conditioned eyeblinks to a mild vibrissal airpuff as the conditioned stimulus while injections of the glutamate antagonist γ-d-glutamylglycine were administered to the IO. Remarkably, even though this treatment suppressed CRs during training sessions, the postacquisition retention test revealed that CR acquisition had not been abolished. The ability to acquire CRs with IO unconditioned stimulus signals that were blocked or severely suppressed suggests that mechanisms responsible for CR acquisition are extremely resilient and probably less dependent on IO-task-related signals than previously thought.